pBACsurf-1

Baculovirus transfer plasmid with the polyhedrin promoter, for expressing GP64 fusion proteins anchored in the membrane of the baculovirus nucleocapsid.

Sequence Author: MilliporeSigma (Novagen)

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PspFI (8770) BseYI (8766) BpmI (8113) NmeAIII (8035) ScaI (7701) EcoO109I (7202) PfoI (7143) DraIII (6786) BanII (6712) SgrDI (5249) SgrAI (4970) BclI * (331) BstXI (1255) baculovirus recombination region (lef2/ORF603) EcoRV (2099) SpeI (2205) BfuAI - BspMI (2310) PstI (2321) Acc65I (2322) KpnI (2326) TspMI - XmaI (2327) SmaI (2329) NruI (2660) SacII (2676) PaeR7I - PspXI - XhoI (2794) BstZ17I (2829) NotI (3106) PflFI - Tth111I (3517) PflMI (3584) SnaBI (3882) SwaI (3972) AgeI (4730) pBACsurf-1 9429 bp
PspFI  (8770)
1 site		 C C C A G C G G G T C G
BseYI  (8766)
1 site		 C C C A G C G G G T C G
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
BpmI  (8113)
1 site		 C T G G A G ( N ) 14 N N G A C C T C ( N ) 14
Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
NmeAIII  (8035)
1 site		 G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19
Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
ScaI  (7701)
1 site		 A G T A C T T C A T G A
EcoO109I  (7202)
1 site		 R G G N C C Y Y C C N G G R
Sticky ends from different EcoO109I sites may not be compatible.
PfoI  (7143)
1 site		 T C C N G G A A G G N C C T
Sticky ends from different PfoI sites may not be compatible.
DraIII  (6786)
1 site		 C A C N N N G T G G T G N N N C A C
Sticky ends from different DraIII sites may not be compatible.
BanII  (6712)
1 site		 G R G C Y C C Y C G R G
Sticky ends from different BanII sites may not be compatible.
SgrDI  (5249)
1 site		 C G T C G A C G G C A G C T G C
SgrAI  (4970)
1 site		 C R C C G G Y G G Y G G C C R C
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
BclI  (331)
1 site		 T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
BstXI  (1255)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
EcoRV  (2099)
1 site		 G A T A T C C T A T A G
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
SpeI  (2205)
1 site		 A C T A G T T G A T C A
BfuAI  (2310)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BfuAI recognition sequence.
Sticky ends from different BfuAI sites may not be compatible.
BfuAI is typically used at 50°C, but is 50% active at 37°C.
BspMI  (2310)
1 site		 A C C T G C ( N ) 4 T G G A C G ( N ) 4 ( N ) 4
Efficient cleavage requires at least two copies of the BspMI recognition sequence.
Sticky ends from different BspMI sites may not be compatible.
PstI  (2321)
1 site		 C T G C A G G A C G T C
Acc65I  (2322)
1 site		 G G T A C C C C A T G G
KpnI  (2326)
1 site		 G G T A C C C C A T G G
TspMI  (2327)
1 site		 C C C G G G G G G C C C
XmaI  (2327)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SmaI  (2329)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
NruI  (2660)
1 site		 T C G C G A A G C G C T
SacII  (2676)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
PaeR7I  (2794)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (2794)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (2794)
1 site		 C T C G A G G A G C T C
BstZ17I  (2829)
1 site		 G T A T A C C A T A T G
NotI  (3106)
1 site		 G C G G C C G C C G C C G G C G
PflFI  (3517)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3517)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
PflMI  (3584)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
Sticky ends from different PflMI sites may not be compatible.
SnaBI  (3882)
1 site		 T A C G T A A T G C A T
SwaI  (3972)
1 site		 A T T T A A A T T A A A T T T A
SwaI is typically used at 25°C, but is 50% active at 37°C.
AgeI  (4730)
1 site		 A C C G G T T G G C C A
GP64 signal sequence
2254 .. 2313  =  60 bp
20 amino acids  =  2.0 kDa
Product: signal sequence from the baculovirus envelope glycoprotein GP64
GP64 signal sequence
2254 .. 2313  =  60 bp
20 amino acids  =  2.0 kDa
Product: signal sequence from the baculovirus envelope glycoprotein GP64
GP64 mature domain
2332 .. 3810  =  1479 bp
492 amino acids  =  56.6 kDa
Product: mature domain of the baculovirus envelope glycoprotein GP64
GP64 mature domain
2332 .. 3810  =  1479 bp
492 amino acids  =  56.6 kDa
Product: mature domain of the baculovirus envelope glycoprotein GP64
baculovirus recombination region (ORF1629)
3819 .. 5176  =  1358 bp
contains part of ORF1629
baculovirus recombination region (ORF1629)
3819 .. 5176  =  1358 bp
contains part of ORF1629
AmpR
7395 .. 8255  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   7395 .. 7463  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7395 .. 8255  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   7464 .. 8255  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
7395 .. 8255  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
baculovirus recombination region (lef2/ORF603)
1269 .. 2096  =  828 bp
contains ORF603 and part of lef2
baculovirus recombination region (lef2/ORF603)
1269 .. 2096  =  828 bp
contains ORF603 and part of lef2
ori
8426 .. 9014  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
8426 .. 9014  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
f1 ori
6553 .. 7008  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
6553 .. 7008  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
AmpR promoter
7290 .. 7394  =  105 bp
AmpR promoter
7290 .. 7394  =  105 bp
polyhedrin promoter
2100 .. 2191  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
2100 .. 2191  =  92 bp
promoter for the baculovirus polyhedrin gene
ORF1629
3835 .. 5176  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF1629
3835 .. 5176  =  1342 bp
446 amino acids  =  49.2 kDa
Product: baculovirus capsid-associated protein
required for viral replication
ORF603
1431 .. 2036  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
ORF603
1431 .. 2036  =  606 bp
201 amino acids  =  23.6 kDa
Product: baculovirus ORF603 protein
lef2
1269 .. 1393  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
lef2
1269 .. 1393  =  125 bp
40 amino acids  =  4.5 kDa
Product: baculovirus late expression factor 2
MCS
2317 .. 2332  =  16 bp
multiple cloning site
MCS
2317 .. 2332  =  16 bp
multiple cloning site
ORF:  451 .. 780  =  330 bp
ORF:  109 amino acids  =  12.4 kDa
ORF:  2200 .. 3810  =  1611 bp
ORF:  536 amino acids  =  61.4 kDa
ORF:  6271 .. 6576  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  761 .. 1393  =  633 bp
ORF:  210 amino acids  =  23.9 kDa
ORF:  5469 .. 6287  =  819 bp
ORF:  272 amino acids  =  32.1 kDa
ORF:  7395 .. 8255  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  9417 .. 248  =  261 bp
ORF:  86 amino acids  =  9.8 kDa
ORF:  2272 .. 3243  =  972 bp
ORF:  323 amino acids  =  36.4 kDa
ORF:  3502 .. 3732  =  231 bp
ORF:  76 amino acids  =  8.0 kDa
ORF:  3835 .. 5088  =  1254 bp
ORF:  417 amino acids  =  46.0 kDa
ORF:  9339 .. 422  =  513 bp
ORF:  170 amino acids  =  19.8 kDa
ORF:  1431 .. 2036  =  606 bp
ORF:  201 amino acids  =  23.6 kDa
ORF:  6216 .. 6533  =  318 bp
ORF:  105 amino acids  =  12.3 kDa
ORF:  1475 .. 1705  =  231 bp
ORF:  76 amino acids  =  9.3 kDa
ORF:  5132 .. 5470  =  339 bp
ORF:  112 amino acids  =  13.3 kDa
ORF:  7859 .. 8125  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Individual Sequences & Maps

BaculoDirect C-Term Linear DNA
pBACgus-5
pIEx Bac-1
pMT BiP V5-His A
BaculoDirect N-Term Linear DNA
pBACgus-6
pIEx Bac-1 Ek_LIC (linearized)
pMT BiP V5-His B
pAB-6xHis-MBP
pBACgus4x-1
pIEx Bac-3
pMT BiP V5-His C
pAB-6xHis
pBacPAK8
pIEx Bac-3 3C_LIC (linearized)
pMT V5-His A
pAB-bee
pBacPAK9
pIEx Bac-4
pMT V5-His B
pAB-bee-8xHis
pBACsurf-1
pIEx Bac-4 Ek_LIC (linearized)
pMT V5-His C
pAB-bee-FH
pBAC-1
pIEx Bac-5
pQE-TriSystem
pAB-GST
pBAC-2cp
pIEx Bac-5 3C_LIC (linearized)
pQE-TriSystem His-Strep1
pAB-MBP
pBAC-3
pIEx-1
pQE-TriSystem His-Strep2
pAc5.1 V5-His A
pBAC-5
pIEx-10
pQE-TriSystem Strep
pAc5.1 V5-His B
pBAC-6
pIEx-10 Ek_LIC
pQE-Trisystem-5
pAc5.1 V5-His C
pBiEx-1
pIEx-10 Ek_LIC (linearized)
pQE-Trisystem-6
pAcAB3
pBiEx-2
pIEx-2
pQE-TriSystem-7
pAcAB4
pBiEx-3
pIEx-3
pTriEx-1.1
pACEBac1
pCoBlast
pIEx-4
pTriEx-1.1 Hygro
pACEBac2
pCoHygro
pIEx-5
pTriEx-1.1 Neo
pAcG2T
pDEST10
pIEx-6
pTriEx-2
pAcG3X
pDEST20
pIEx-7 Ek_LIC
pTriEx-2 Hygro
pAcGHLT A
pDEST8
pIEx-7 Ek_LIC (linearized)
pTriEx-2 Neo
pAcGHLT B
pDream2.1 MCS
pIEx-8
pTriEx-3
pAcGHLT C
pFastBac Dual
pIEx-8 Ek_LIC
pTriEx-3 Hygro
pAcGP67-A
pFastBac CT-TOPO (linearized)
pIEx-8 Ek_LIC (linearized)
pTriEx-3 Neo
pAcGP67-B
pFastBac HBM-TOPO (linearized)
pIEx-9
pTriEx-4
pAcGP67-C
pFastBac NT-TOPO (linearized)
pIEx-9 3C_LIC
pTriEx-4 Ek_LIC (linearized)
pAcHLT A
pFastBac1
pIEx-9 3C_LIC (linearized)
pTriEx-4 Hygro
pAcHLT B
pFastBacHT A
pIZ V5-His
pTriEx-4 Neo
pAcHLT C
pFastBacHT B
pIZT V5-His
pTriEx-5
pAcMP2
pFastBacHT C
pMelBac A
pTriEx-5 Ek_LIC (linearized)
pAcMP3
pI-secSUMOstar
pMelBac B
pTriEx-6
pAcSecG2T
pI-SUMOstar
pMelBac C
pTriEx-6 3C_LIC (linearized)
pAcSG2
pIB V5-His
pMIB V5-His A
pTriEx-7
pAcUW51
pIB V5-His-DEST
pMIB V5-His B
pTriEx-7 Ek_LIC (linearized)
pBAC4x-1
pIDC
pMIB V5-His C
pVL-FH
pBACgus-1
pIDK
pMT-DEST48
pVL1392
pBACgus-2cp
pIDS
pMT BioEase-DEST
pVL1393
pBACgus-3

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